21 August 2006
Introduction to Proteins I
Dr. Kandice Williams, Ph.D.



Covalent Bonds


  • Strongest biochemical bond
    • Example: benzene resonance structure
    • Peptide bonds take 732 kJ/mol to dissociate
  • Formed by sharing a pair of electrons between adjacent atoms
  • Chemical reactions requiring the most energy break and form covalent bonds

Electrostatic Interactions


  • Depend on electrical charge of atoms
    • Atoms with single opposite charges in water take ~6 kJ/mol to dissociate
    • Dielectric constant of water is very high, D = 80

Hydrogen Bonds


  • Interactions are also weak electrostatic interactions
    • Longer than covalent bonds
  • Energy of dissociation = ~4-13 kJ.mol
  • Hydrogen atom is shared between two electron negative atoms (nitrogen or oxygen)
    • One is the hydrogen bond donor (+), the other is the hydrogen bond acceptor (-)

van der Waal Interactions


  • Important in hydrophobic environments such as the interior of a molecule
  • depend on the distance between two atoms and non-symmetrical charge distribution around each
  • van der Waals contact distance is the point at which two atoms exhibit greatest attraction for each other
    • Too close and atoms repulse each other, but too far and the interaction is negligable
    • Contact point is approximately 2-3 Å
    • Energy of dissociation = ~2-4 kJ/mol

Water


  • Water affects non-covalent bonds which are important for biochemical reactions
    • Non-covalent interactions are important for biochemical reactions because they are easily reversible
  • Water is polar with asymmetric distribution of charge and highly cohesive through hydrogen bonding
  • High boiling point, heat of vaporization, heat of fusion, surface tension, internal cohesion, and dielectric constant
  • Excellent solvent for polar molecules

Hydrophilic Effect

  • High dielectric constant diminishes strength of electrostatic attractions between other polar molecules
  • Forms solvent (hydrated) shells around other polar molecules, creating new electrostatic fields
  • Rapid fluctuating hydrogen bond structure that allows other molecules to diffuse and interact
  • Allow high concentrations of other polar molecules to exist in water as a solution.

Hydrophobic Effect

  • Water interaction with non-polar molecules
  • Non-polar molecules aggregate together in water following the second law of thermodynamics to increase the total entropy of the system and release free energy
    • Hydrophobic effect promotes many biochemical reactions such as correct protein folding
      • Hydrophilic amino acids move towards the exterior
      • Hydrophobic amino acids move to the interior of the protein

Acids and Bases


  • Acid – H+ Donor
    • Strong acid produces a weak conjugate base – poor pH buffering
    • Weak acid produces a strong conjugate base – good pH buffering
  • Base – H+ Receiver
    • Strong base produces a weak conjugate acid – poor pH buffering
    • Weak base produces a strong conjugate acid – good pH buffering
  • Ka = Acid Equilibrium Constant
    • Ka = [H+][A-] / [HA]
    • [H+] remains constant when pH buffering is strong

pH

  • Logarithmic measure of the concentration of [H+]
  • pH = -log[H+]
    • Pure H2O at room temperature: [H+] equals [OH-] = 1 x 10-7 M
      • Therefore, -log[H+] = 7 and the pH of water is 7

pK

  • pK is the pH when the [conjugate acid] = [conjugate base]
  • Resistance to pH change is greatest (greatest buffering capacity)
    • pKa of an acid is the pH when [HA] = [A-]

Henderson-Hasselbalch Equation

  • Predicts the pH of a buffer by the –log [HA] / [A-]
  • For a weak acid, pH = pKa + log [A-] / [HA]
  • When [A-] equals [HA], the pH = pKa

Properties of a Buffer

  • Acid-base conjucate pair resists changes in pH of solution
  • Weak acid and strong conjugate base make a strong pH buffer
    • Bicarbonate buffer system in blood
      • Kidneys regulate H+ by renal excretion while lungs regulate CO2 by rate of ventilation
      • H+ + HCO3- <--> H2CO3 <--> H2O + CO2

Amino Acids



  • All proteins use the same set of 20 amino acids for the last several billion years
  • Amino acids vary in size, shape, charge, and chemical reactivity
  • Zwitterions (hybrid ions):
    • Amino acids in solution at physiological pH exist predominately as dipolar ions
      • Amine group is protonated (-NH3+)
      • Carboxyl group is deprotonated (-COO-)
      • Therefore, amino acid is fully ionized but electrically neutral
      • Amino acids are conjugate acid/base pairs (can be diprotic or tripotic depending on –R group)
  • Amino acid –R groups facilitate chemical reactions and form ionic bonds
    • All three pKa’s of the –COOH, -NH3+, and –R group determine the pI
  • pI is the isoelectric point which is the pH when the total net charge is zero
    • Amino acids at their isoelectric point will not move in an electric field.
  • Amino Acids have 4 different groups, (1) -NH3+, (2) –COO-, (3) –R group, (4) Hydrogen
    • Amino Acids are chiral and L-α-amino acids are constituents of proteins
      • Have an S (left) chiral configuration.


  • This is a Powerpoint slideshow with the amino acid structures (like flashcards) that Habib made in undergrad. Thought it might help someone:


Non-polar, aliphatic –R groups

  • Hydrophobic – the larger the aliphatic side chain, the more hydrophobic
  • Glycine
    • Non-chiral because –R group is another hydrogen
  • Alanine
  • Valine
  • Leucine
  • Isoleucine
    • contains an additional chiral center in its –R group
  • Methionine
    • contains a thioether (-S-) group and described as “polar uncharged”
  • Proline
    • Aliphatic –R group is bonded to both nitrogen and α-carbon, making it conformationally restricted

Aromatic –R groups

  • Generally non-polar because of aromatic rings
  • Phenylalanine
  • Tyrosine
    • Have hydrophilic properties because of –OH group
    • Strongly absorb UV light near 280 nm, which can be used to estimate the concentration of proteins in a solution
  • Tryptophan
    • Have hydrophilic properties because of –NH- group
    • Also strongly absorb UV light near 280 nm

Aliphatic hydroxyl –R groups

  • Hydrophilic attributes due to –OH group and stronger chemical reactivity
  • Serine
    • Hydroxylated version of alanine
  • Threonine
    • Like valine but with a –OH group replacing –CH3

Aliphatic sulfhydral (thiol) –R group

  • Polar amino acid
  • Cysteine
    • -SH groups from two cysteine amino acids can form a covalently-linked cystine disulfide bridge
      • Requires an oxidation reaction
    • Similar to serine but contains a more reactive –SH group

Basic –R groups

  • Very polar and hydrophilic
  • -R groups are positively charged a physiological pH
  • Lysine
  • Arginine
  • Histidine
    • Most reactive because it accepts H+

Acidic –R groups and Uncharged derivatives

  • Acidic side chains that are usually negatively charged at physiological pH
  • H+ Donors
  • Aspartate
  • Glutamate
  • Asparagine
    • Uncharged derivative of Aspartate; contains a terminal carboxamide instead of carboxylic acid
  • Glutamine
    • Uncharged derivative of Glutamate; contains a terminal carboxamide instead of carboxylic acid

Non-Essential and Essential Amino Acids


Objectives